gel electrophoresis by suresh b i
Download as PPTX, PDF4 likes1,625 views
Gel electrophoresis is a method used to separate molecules like proteins and nucleic acids based on their physical properties. It works by applying an electric field to move charged molecules through a gel medium. Key aspects include applying samples to the gel, running electrophoresis with an electric current, and visualizing separated molecules by staining and quantification using a densitometer. Gel electrophoresis has applications in forensics, genetics, and biochemistry to analyze DNA, diagnose diseases, and study proteins. It separates biomolecules based on factors like size, shape, and charge.
More Related Content
What's hot (20)
Similar to gel electrophoresis by suresh b i (20)
Recently uploaded (20)
gel electrophoresis by suresh b i
- 1. WELCOME 1
- 2. GEL ELECTROPHORESIS Guided by: Ms. Ashwini D Dept. of Industrial chemistry Presented by: Suresh B I 2
- 3. ELECTROPHORESIS Electrophoresis is the migration of charged particles in an electric field towards the electrode bearing the opposite charge. (OR) Is a method where by charged molecules in solution, like proteins and nucleic acids migrate in response to an electric field 3
- 4. PRINCIPLE Electrophoresis is the process of migration of charged molecules through solution in an applied electric field. Electrophoresis is often classified according to the presence or absence of a solid supporting medium or matrix through which the charged molecules moves in the electrophoretic system. Solution electrophoresis system employs aqueous buffers in the absence of solid support medium. Such system can suffer from sample mixing due to diffusion of the charged molecules with resolution loss of resolution during the sample application separation & removal steps. 4
- 5. Separation of molecules on the basis of >size and/or >charge and/or >shape 5
- 6. 6
- 7. TYPES OF ELECTROPHORESIS Based on supporting media (zone electrophoresis): Paper electrophoresis gel electrophoresis Cellulose acetate electrophoresis Without supporting media (moving boundary method): Free electrophoresis Supporting medias are : starch, Agar- Agarose, polyacrylamide , 7
- 9. 9
- 10. GEL ELECTROPHORESIS Introduction: Gel electrophoresis is group of techniques used by scientists to separate molecules based on physical characteristics such as size, shape, and isoelectric point. Gel electrophoresis is usually performed for analytical purposes. 10
- 11. APPICATION OF SAMPLE RUNNING OF SAMPLE VISUALISATION OF SAMPLE QUANTIFICATION TECHNIQUES 11
- 12. REQUIREMENTS FOR GEL ELECTROPHORESIS Buffer Fixative Staining solution Destaining solution Densitometer Is essentially a double beam filter photometer or spectrophotometer that scans the electrophoretic strip ( in the form of agarose , cellulose acetate or polyacrylamide) as it moves past the optical system. 12
- 13. DENSO METER 13
- 14. PROCEDURE 1. Sample to be separated is applied to a supporting medium (paper, cellulose acetate, agar gel, polyacrylamide gel etc.) Electrophoresis is carried out at desired constant voltage or constant current in presence of specific pH. 3. After completion of electrophoresis the supporting medium is placed in a fixative to prevent diffusion of separated fractions. 4. Separated fraction is then visualized by using appropriate stains e.g. Bromophenol Blue & Amino Schwartz for plasma proteins and Sudan Black for lipoproteins. 5. Quantification of each fraction is done by either densitometer or elution followed by colorimeter or spectrophotometer of eluted fraction. 14
- 16. Gel electrophoresis is a process that separates fragments of DNA based on their sizes. 16
- 17. APPLICATION OF GEL ELECTROPHORESIS Evidence in criminal cases To determine paternity To diagnose genetic diseases Help to determine kinship in animals Compare similarities and differences between species Solve paternity cases Determine genetic kinship among species 17
- 18. Estimated molecular masses and relative abundance of unknown polypeptides in a complex mixture Patterns of bands that suggest presence of isoenzymes or specific complex proteins Effectiveness of a separation procedure during cell/tissue fractionation Effectiveness of a procedure to purify specific organelles, proteins, or polypeptides 18
- 19. CONCLUSION Gel electrophoresis is used in forensic, molecular biology and bio chemistry The results can be analysed quantitatively by visualizing the gel with UV - light and a gel imaging device The image is recorded with a computer operated camera and the intensity of the band (or) spot of interest is measuring and compared against standard (or)markers loaded on the sample gel The measurement and analysis are mostly done with specialised software's 19
- 20. REFERENCE colloidal chemistry by G Whitmore published by prabhath Kumar Sharma or sarup and sons page no:44 colloidal chemistry by B K Sharma page no:84 20
- 21. 21